Capsid variants and methods of using the same

Abstract

The disclosure is directed in part to variant capsid polypeptides that can be used to deliver payloads.

1 - 49 . (canceled) 
     
     
         50 . A variant adeno-associated virus (AAV) capsid polypeptide having at least 95% identity to the VP1, VP2, or VP3 sequence of SEQ ID NO: 13, comprising an insertion of IEHWGH (SEQ ID NO: 43) between positions 450 and 451 according to SEQ ID NO: 1. 
     
     
         51 . The variant AAV capsid polypeptide of  claim 50 , which has at least 96% sequence identity to the VP1, VP2, or VP3 sequence of SEQ ID NO: 13. 
     
     
         52 . The variant AAV capsid polypeptide of  claim 50 , which has at least 97% sequence identity to the VP1, VP2, or VP3 sequence of SEQ ID NO: 13. 
     
     
         53 . The variant AAV capsid polypeptide of  claim 50 , which has at least 98% sequence identity to the VP1, VP2, or VP3 sequence of SEQ ID NO: 13. 
     
     
         54 . The variant AAV capsid polypeptide of  claim 50 , which has at least 99% sequence identity to the VP1, VP2, or VP3 sequence of SEQ ID NO: 13. 
     
     
         55 . A nucleic acid molecule encoding the variant AAV capsid polypeptide of  claim 50 . 
     
     
         56 . A variant AAV capsid polypeptide comprising the VP1, VP2 or VP3 sequence of SEQ ID NO: 13. 
     
     
         57 . A nucleic acid molecule encoding the variant AAV capsid polypeptide of  claim 56 . 
     
     
         58 . The nucleic acid molecule of  claim 57  comprising SEQ ID NO: 31. 
     
     
         59 . A recombinant AAV virus particle comprising the AAV variant capsid polypeptide of  claim 50 . 
     
     
         60 . The recombinant AAV virus particle of  claim 59 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene. 
     
     
         61 . A recombinant AAV virus particle comprising the variant AAV capsid polypeptide of  claim 56 . 
     
     
         62 . The recombinant AAV virus particle of  claim 61 , comprising a nucleic acid molecule comprising a promoter operably linked to a heterologous transgene. 
     
     
         63 . A host cell comprising the AAV variant capsid polypeptide of  claim 50 . 
     
     
         64 . A host cell comprising the nucleic acid molecule of  claim 55 . 
     
     
         65 . A host cell comprising the AAV variant capsid polypeptide of  claim 56 . 
     
     
         66 . A host cell comprising the nucleic acid molecule of  claim 57 . 
     
     
         67 . A method of delivering a payload to a cell comprising contacting the cell with the recombinant AAV virus particle of  claim 59 . 
     
     
         68 . A method of producing an AAV virus particle comprising AAV capsid polypeptides, the method comprising:
 introducing a nucleic acid molecule of  claim 55  into a cell,   culturing the cell under conditions to produce the AAV virus particle; and   harvesting the AAV virus particle.   
     
     
         69 . A recombinant AAV comprising:
 (a) an AAV capsid comprising AAV capsid polypeptides, each having an amino acid sequence comprising the VP1, VP2 or VP3 sequence of SEQ ID NO: 13; and   (b) a minigene having AAV inverted terminal repeats and a transgene comprising a heterologous gene operably linked to regulatory sequences which direct expression of the heterologous gene in a host cell.